Sea island cotton receptor analogous protein kinase gene and its application

A technology of protein kinase gene and receptor, which is applied in the field of cloning of receptor-like protein kinase gene of sea island cotton, to achieve considerable economic benefits, reduce losses, and improve the effect of verticillium wilt resistance

Inactive Publication Date: 2007-06-06
NANJING AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] There are 14 cotton kinase-related sequences registered in Genbank, among which the known functions are all involved in the development of cotton fiber, while cotton kinase genes related to disease resistance have not been reported yet

Method used

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  • Sea island cotton receptor analogous protein kinase gene and its application
  • Sea island cotton receptor analogous protein kinase gene and its application

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Experimental program
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Effect test

Embodiment Construction

[0063] 1. Design degenerate primers using the conserved domains of kinase-like disease-resistant genes that have been cloned in other crops, and perform PCR amplification from Haidao Mianhai 7124. Fragments of expected size were recovered, and the recovered fragments were ligated into pGEM-T easy Vector (Promega). Pick 10 positive clones and send them to Shanghai Handsome Biotechnology Co., Ltd. for sequencing.

[0064] 2. Sequencing results After manual removal of vector sequences by DNAstar software, BLASTx analysis was performed in the non-redundant protein database of NCBI.

[0065] 3. The 7 sequences related to kinases in the BLASTx analysis results were analyzed with DNAstar for continuous open reading frames (ORFs), and 3 sequences with continuous ORFs were found (sequence numbers: L354, L355, L361, respectively).

[0066] 4. Design specific primers using 3 sequences with continuous ORFs. Analysis of its expression in different periods in root tissues of pirate cotton...

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Abstract

The present invention relates to the cloning of sea island cotton receptor analogous protein kinase gene, and belongs to the field of biotechnology. The present invention clones the full length cDNA sequence of sea island cotton receptor analogous protein kinase gene, designs one pair of specific primers for sea island cotton receptor analogous protein kinase, detects the expression of the gene in resistant sea island cotton breed Hai-7124 with inoculated verticillum wilt in the RT-PCR method, and verifies the result by means of Northern hybridization. It is shown that the gene is expressed only in 96 hr after verticillum wilt treatment, so that the gene is relate directly with verticillum wilt. The gene may be used as the target gene in culturing verticillum wilt resisting cotton breed.

Description

(1) Technical field [0001] The invention relates to the cloning of a receptor-like protein kinase gene of Gossypia sea-island cotton and belongs to the field of biotechnology. (2) Technical background [0002] Cotton is an important economic crop, and cotton fiber is the main textile raw material. However, there are many diseases of cotton, and the blight and verticillium wilt are particularly serious in my country, causing great losses to cotton production. Planting disease-resistant varieties is the most cost-effective way to control the disease. Upland cotton is the main variety of cotton planted in my country. Upland cotton breeding for resistance to Fusarium wilt has been very successful, but the cultivation of Verticillium wilt-resistant varieties has always been a major problem for breeders. The main reason is that there is no immunity or high resistance in upland cotton. Verticillium wilt resistance source. [0003] Cotton verticillium wilt was discovered and repor...

Claims

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Application Information

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IPC IPC(8): C12N15/54C12N9/12
Inventor 张天真高玉龙郭旺珍
Owner NANJING AGRICULTURAL UNIVERSITY
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